Nanoparticles for the administration of active ingredients, method of producing said particles and composition containing same

ABSTRACT

The invention relates to nanoparticles used for the administration of active ingredients, the method of producing said nanoparticles and compositions containing same. The inventive nanoparticles comprise a chitosan, glucomannan, at least one active ingredient and, if necessary, an anionic salt, preferably in sodium tripolyphosphate form. Said nanoparticles can be used to administer active ingredients to a human or animal body.

FIELD OF THE INVENTION

The present invention refers to nanoparticles comprising chitosan,glucomannan, at least one active ingredient and, if necessary, ananionic salt, preferably in sodium tripolyphosphate form, which can beused to administer active ingredients to the human or animal body. Itfurther refers to a method for obtaining said nanoparticles, and to acomposition comprising said nanoparticles.

BACKGROUND OF THE INVENTION

It is known that the administration of a number of active ingredients bydifferent administration routes to the human or animal body has variousdifficulties. It is especially worth indicating the difficulties ofadministration by mucosal routes, especially peptides and proteins,given that said administration is strongly affected by the limitedpermeability of the epithelial barriers of the human or animal body. Itis also known that it is possible to overcome part of these difficultiesby incorporating the active ingredients which are to be administered insmall particles. The transport of said particles through the mucosae isaffected mainly by the size of these particles, the transport increasingwith the decrease of the particle size. Therefore, the transport of, forexample, nanoparticles (generally, with a mean diameter of less than 1μm), through mucosae is better than that of microparticles (generally,with a mean diameter of 1 μm up to several hundred elm). In fact, thetransport of nanoparticles through the mucosae of the human or animalbody occurs naturally. It is also known that the effectiveness of theinteraction between the nanoparticles with the epithelial cells can beimproved by means of the incorporation of the nanoparticles of materialscontaining specific ligands. For example, the absorption of liposomes byM cells can be improved by coating said liposomes with mannan residues,as disclosed in the documents Takada et al., Biochim. Biophys. Acta 802,1984, 237-243 and Tomizawa et al., Pharm. Res. 10, 1993, 549-552. It isfurther known that nanoparticles constituted of chitosan foment thetransport of macromolecules incorporated therein through the nasal andintestinal epithelium.

A series of publications are known in the State of the Art referring tochitosan nanoparticles for the administration of active ingredients andto methods for obtaining said nanoparticles. Among said publications, itis worth pointing out Calvo et al., J. Appl. Polym. Sci., 1997, 63,125-132; Calvo et al., 14, 1997b, 1431-1436; Fernández-Urrusuno et al.,Phar. Res. 16, 1991a, 1576-1581; Fernández-Urrusuno et al., S. T. P.Pharm. Sci. 9, 1999b, 429-436; Tokumitsu et al., Pharm. Res. 16, 1999,1830-1835; Mitra et al., J. Control. Release 74, 2001, 317-323; U.S.Pat. No. 2,001,051189 and U.S. Pat. No. 5,843,509. All thesenanoparticies, the main component of which is chitosan, have thedrawback of not being stable at given pH values, specifically, theydissolve in an acid medium and precipitate in a basic medium.

Patent document WO-A-01/01964 for its part refers to compositions forthe sustained release of biomolecules, in microparticle form, comprisingan anionic polymer and another cationic polymer, which interact with oneanother, and biomolecules. The cationic polymer can be a water soluble,positively charged polymer, such as chitosan, for example. Dextransulfate, heparin, alginic acid, alginate, carrageenan, an anionicpolymethacrylate and a positively charged polyamino acid are mentionedas anionic polymers.

Patent document WO-A-96/20698 refers to nanoparticles for the sustainedrelease of bioactive agents, comprising a core based on a biocompatibleand biodegradable polymer, which can be chitosan, said nanoparticleshaving associated or incorporated at least one bioactive agent and atleast one surface modifying agent. It also refers to a method forobtaining the nanoparticles, based on the mixture of organic solutionsof the components, and their subsequent addition to an aqueous phase,with subsequent evaporation of the organic solvent and separation of thenanoparticles from the resulting aqueous phase.

Patent document WO-A-01/32751 refers to a method for producing chitosansor chitosan derivatives in nanoparticle form, with a mean particlediameter in the range of 10 to 1000 nm, consisting of dissolving thechitosan or chitosan derivative in an aqueous acid medium and raisingthe pH of the solution in the presence of a surface modifying agent,until the precipitation of the chitosan is achieved.

Patent document WO-A-99147130 refers to nanoparticles having abiocompatible and biodegradable polyelectrolyte complex, starting fromat least one polycation (which can be chitosan) and at least onepolyanion, as well as at least one bioactive ingredient, thenanoparticles being obtainable by additionally treating thepolyelectrolyte complex during or after its formation with at least onecross-linking agent (glyoxal, TSTU or EDAP).

Most known nanoparticle and microparticle systems produced on the basisof chitosan have the important drawback of being unstable after theiradministration in vivo, as well as during the storage thereof. There areusually difficulties in the lyophilization process, specificallydifficulties in the reconstitution of the lyophilized systems, whichrepresents an important additional limitation for the suitableexploitation of this type of systems. In addition, for thelyophilization of these systems, disclosed in the state of the art, itis necessary to add high amounts of sugars. As a result of theforegoing, the known nanoparticles and microparticles must generally bestored in a liquid suspension form; this usually results in thedestruction of these systems in a few months.

For its part, glucomannan has traditionally been used as a dietsupplement, for the purpose of reducing cholesterol level, in additionto the fact that it has been used in cosmetic applications. The use ofglucomannan in the preparation of pharmaceutical compositions isdisclosed in a series of documents. For example, its use in thepreparation of pharmaceutical compositions in gel form is mentioned inWO-A-99/01166, and U.S. Pat. No. 5,662,840; Xiao et al., J. Appl. Polym.Sci. 76, 2000, 509-515 and U.S. Pat. No. 6,159,504 mention their use inpharmaceutical compositions in film form; US-A-2002019447 and U.S. Pat.No. 2,002,018812 mention pharmaceutical compositions in foam, capsuleand sponge form; and U.S. Pat. No. 6,221,393 in compositions in tabletform. Some of these documents mention the possible incorporation ofchitosan to the pharmaceutical compositions.

It has been mentioned in some documents the possible existence of aninteraction between chitosan and glucomannan, for example in film form,in Xiao et al., J. Appl. Polym. Sci. 76, 2000, 509-515, as well as ingranule form with a diameter exceeding 1 mm, for administering analgesicdrugs, in Xie et al., J. Macromol. Sci., Pure Appl. Chem. A29, 1992,931-8 and Xie et al., J. Clin. Pharm. Sci. 1, 1992, 42-8. Germanpublication DE19839515 refers to a pharmaceutical preparation containingat least one polymer-colloidal active ingredient (particle size <1 μm)association product, in which at least one component is a biocompatibleand biodegradable polymer, which can be chitosan.

In accordance with all the foregoing, there is a need to provide a typeof nanoparticles with improved properties of absorption by the human oranimal body, especially through the mucosal tissue, and which could bestored for a longer time period without undergoing importantalterations. Furthermore, the methods for preparing nanoparticles,whether based on chitosan or not, usually require the use of organicsolvents, which have the drawbacks known by any person skilled in theart, such as their toxicity and the difficulty of eliminating them fromthe nanoparticles, whereby it is appropriate to find a method forproducing nanoparticles complying with the above-mentioned properties,which is simple and does not require the use of organic solvents.

DESCRIPTION OF THE INVENTION

It has now been found that nanoparticles comprising chitosan,glucomannan, the active ingredient to be administered and, optionally,an anionic salt, preferably sodium tripolyphosphate, solve the mentioneddrawbacks of the state of the art. A simple method of preparing theprevious nanoparticles, without the need of using organic solvents, hasfurther been found.

According to a first aspect, the present invention refers to a methodfor producing nanoparticles with a mean diameter of less than or equalto 1 μm, incorporating at least one active ingredient, comprising thefollowing steps:

a) preparing an aqueous chitosan solution,

b) preparing an aqueous glucomannan solution, and

c) mixing, under stirring, the solutions of steps a) and b), such thatthe chitosan and glucomannan nanoparticles are obtained,

wherein at least one of the solutions of steps a) and b) contains atleast one active ingredient.

According to a second aspect, the present invention refers tonanoparticles obtained according to the previous method, comprisingchitosan, glucomannan and at least one active ingredient.

According to an additional aspect, the invention refers to apharmaceutical or cosmetic composition comprising the previousnanoparticles, together with at least one pharmaceutically orcosmetically acceptable excipient, respectively.

According to a preferred embodiment of the method, the glucomannansolution further contains an anionic salt, preferably intripolyphosphate sodium form, for the purpose of favoring thespontaneous formation of the nanoparticles.

Preferably, the concentration of the chitosan solution used in themethod is in the range between 0.5 and 5 mg/mL.

Also preferably, the concentration of the glucomannan solution of themethod is in the range between 0.1 and 50 mg/mL.

The ratio of chitosan with regard to glucomannan (by weight) may vary,preferably between 1:0.02 to 1:100, particularly preferred between 1:0.5and 1:50.

During the method of producing the nanoparticles, the pH value of thechitosan solution is preferably maintained at a pH between 2 and 6.

The method of producing the chitosan and glucomannan nanoparticles canfurther comprise an additional step, in which said nanoparticles arelyophilized. Unlike in the nanoparticles known in the state of the art,for the lyophilization of the nanoparticles according to the presentinvention, only the addition of small amounts of sugars is needed,although it is also possible to carry out the lyophilization withoutadding sugars. In their lyophilized form, the nanoparticles can bestored for longer periods of time, and they can be easily regenerated,when needed, by means of adding the necessary amount of water.

According to this additional embodiment in which the obtainednanoparticles are lyophilized, the present invention further refers tothe chitosan and glucomannan nanoparticles, according to the invention,lyophilized, and to a pharmaceutical or cosmetic composition comprisingsaid lyophilized nanoparticles, and at least one pharmaceutically orcosmetically acceptable excipient.

The chitosan and glucomannan nanoparticles obtainable by means of themethod described above have better stability upon contact withbiological fluids and also during storage than the nanoparticles knownin the state of the art. In fact, the chitosan and glucomannannanoparticles are stable in both an aqueous acid and basic medium,whereby they can be stored in liquid suspension form for long periods oftime. They also have an improved capacity of absorption by the human oranimal body.

Additionally, the chitosan and glucomannan nanoparticles are systems ofboth pharmaceutical and cosmetic utility. They can further beadministered by several routes, such as, for example, the topical, oral,nasal, pulmonary, vaginal and subcutaneous routes. The active ingredientto be incorporated in the chitosan and glucomannan nanoparticles is theingredient for which the formulation is intended. This ingredient willhave an effect on the human or animal organism after its administration;said effect may cure, minimize or prevent a disease. The activeingredient can be a drug, a vitamin, a vaccine, etc., or a cosmeticagent, intended for improving the physical and aesthetic appearance (forexample, skin moisturizing).

The chitosan and glucomannan nanoparticles according to the presentinvention have a high capacity of association of bioactivemacromolecules, for example insulin, bovine serum albumin or immunogenicproteins. The capacity of association depends on the type ofmacromolecule incorporated. Likewise, for certain macromolecules, thedegree of association may depend on the deacetylation degree of thechitosan: the higher the deacetylation degree, the greater theeffectiveness of association.

However, it is also possible to incorporate other active ingredients tothe nanoparticles, both of a lipophilic and hydrophilic nature. It ispossible to point out, for example, the effective incorporation ofindomethacin (moderately lipophilic) and acyclovir (hydrophilic).

The active ingredient to be incorporated in the nanoparticles isdissolved in one of the two aqueous solutions used in the formation ofthe nanoparticles. In the case of lipophilic active ingredients, thesemust be previously dissolved in a polar organic solvent, miscible withaqueous media, and then it will be added to the chitosan solution, or tothe glucomannan solution. In the case of incorporating more than oneactive ingredient to the nanoparticles, these can be dissolved either inthe same solution or in different solutions.

In the case of incorporating bioactive macromolecules, these canpreferably be incorporated according to the following methods:

-   i) the macromolecule is dissolved in sodium tripolyphosphate, and    the resulting mixture is incorporated to the glucomannan solution    used for producing the nanoparticles;-   ii) the macromolecule is dissolved in a NaOH solution; the obtained    solution is added to sodium tripolyphosphate; the resulting mixture    is incorporated to the glucomannan solution used for producing the    nanoparticles;-   iii) the macromolecule is dissolved in sodium phosphate at pH 6.6;    the obtained solution is added to sodium tripolyphosphate; the    resulting mixture is incorporated to the glucomannan solution used    for producing the nanoparticles;-   iv) the macromolecule is added to the chitosan solution used for    producing the nanoparticles;-   v) the macromolecule is added to a NaOH solution; the obtained    mixture is added to the chitosan solution used for producing the    nanoparticles;-   vi) the macromolecule is added to a sodium phosphate solution at pH    6.6; the obtained mixture is added to the chitosan solution used for    producing the nanoparticles.

The chitosan and glucomannan nanoparticles are colloidal, i.e. theirmean diameter is equal to or less than 1 μm. The mean particle size ismainly affected by the ratio of chitosan with regard to glucomannan, bythe deacetylation degree of the chitosan, by the incorporation of ananionic salt, for example sodium tripolyphosphate, and by the nature ofthe active ingredient. On the other hand, the nanoparticles can have apositive or negative surface charge (measured by means of the zetapotential), the magnitude of which depends on the composition of thenanoparticles and the deacetylation degree of the chitosan.

The nanoparticles further have the capacity to release the activeingredient incorporated therein in a sustained and/or controlled manner.The release of the active ingredient can be controlled by combiningfactors such as the ratio of chitosan with regard to glucomannan, thedegree of acetylation of the chitosan and the method of producing thenanoparticles.

Other purposes, features and advantages of the invention will becomeclearer below in light of the explanatory description which follows,made in reference to several illustrative examples which by no meansimply any limit to the scope of the invention.

EXAMPLES

Through out the examples, the following abbreviations will be sued:

CS=Chitosan

GM=Phosphorylated glucomannan

TPP=Sodium tripolyphosphate

P1: Plant origin protein (Ricinus communis) constituted of twopolypeptides.

Example 1

Chitosan (with an 88% deacetylation degree), glucomannan and sodiumtripolyphosphate nanoparticles were prepared according to the method ofthe invention, with different glucomannan ratios. Once they wereprepared, their mean diameter and zeta potential were measured. TABLE 1CS/TPP/GM Mean diameter Zeta potential (w/w) (nm) (mV) 6/1/2.3 250 ± 24+32.2 ± 2.0 6/1/4.6 302 ± 26 +15.2 ± 1.7

Example 2

Chitosan (with an 88% deacetylation degree) and glucomannannanoparticles were prepared according to the method of the invention,with different glucomannan ratios and without adding any anionic salt.Once they were prepared, their mean diameter and Z potential weremeasured. TABLE 2 CS/GM Mean diameter Zeta potential (w/w) (nm) (mV)6/4.6 252.1 ± 15 +31.25 ± 1.06 6/13.8 185.5 ± 3  +33.2 ± 0.8

Example 3

Chitosan (with an 88% deacetylation degree) and glucomannannanoparticles were prepared, incorporating sodium tripolyphosphate,according to the method of the invention, with different chitosan andglucomannan ratios, incorporating the P1 protein or insulin. Once theywere prepared, their mean diameter and zeta potential were measured.TABLE 3 CS/TPP/GM Associated CS/protein Mean diameter Zeta potential(w/w/w) protein (w/w) (nm) (mV) 4/1/1.5 P1 1.6/1 552 ± 4 +32.1 ± 1  6/1/4.6 P1 1.3/1 296 ± 6 +15.9 ± 0.2 6/1/4.6 P1 2.2/1 263 ± 6 +30.3 ±0.6 6/0.7/4.6 Insulin 2/1 265.8 ± 6   +32.2 ± 0.4

Example 4

Chitosan (with an 88% deacetylation degree) and glucomannannanoparticles were prepared, without incorporating any anionic salt,according to the method of the invention, with different chitosan andglucomannan ratios, incorporating the P1 protein or insulin. Once theywere prepared, their mean diameter and zeta potential were measured.TABLE 4 CS/GM Associated Mean diameter Zeta potential (w/w) protein (nm)(mV) 6/4.6 Insulin 252.3 ± 4 +15.5 ± .06 6/4.6 P1   205 ± 11  +8.5 ± 2.76/13.8 P1 293.2 ± 5 +11.9 ± 3.2

Example 5

Chitosan (88% deacetylation degree) and glucomannan nanoparticles wereprepared, incorporating sodium tripolyphosphate, according to the methodof the invention, with a CS/TPP ratio of 3:1, incorporating the P1protein (25% theoretical load) therein, according to different methods.The effectiveness of association of the P1 protein to the nanoparticles,as well as the load capacity thereof, were measured. TABLE 5Effectiveness of association Load capacity Method (%) (%) TPP^(i) 15.4 ±2.7 8.1 ± 1.4 NaOH-TPP^(ii)  5.6 ± 0.3 3.2 ± 0.2 Phosphate-TPP^(iii)22.6 ± 0.2 11.1 ± 0.1  CS^(iv) 15.5 ± 2.7 7.1 ± 1.2 NaOH—CS^(v)  8.7 ±3.2 5.1 ± 1.8 Phosphate-CS^(vi) 26.0 ± 0.1 11.9 ± 0.5 P1 was dissolved in^(i)TPP;^(ii)NaOH, and added to TPP^(iii)sodium phosphate at pH 6.6, and added to TPP^(iv)CS;^(v)NaOH, and added to CS; and^(vi)sodium phosphate at pH 6.6, and added to CS

Example 6

Chitosan (88% deacetylation degree) and glucomannan nanoparticles wereprepared, incorporating sodium tripolyphosphate, according to the methodof the invention, with a CS/TPP ratio of 6:1, incorporating P1 protein(25% theoretical load) therein, according to different methods. Theeffectiveness of association of the P1 protein to the nanoparticles, aswell as the load capacity thereof, were measured. TABLE 6 Effectivenessof association Load capacity Method (%) (%) TPP^(i) 15.4 ± 4.0 16.8 ±4.9 NaOH-TPP^(ii)  8.7 ± 1.6 10.5 ± 1.9 Phosphate-TPP^(iii) 26.3 ± 1.227.6 ± 1.3 CS^(iv)  6.5 ± 1.7  9.8 ± 2.6 NaOH—CS^(iv) 18.2 ± 1.9 21.4 ±2.4 Phosphate-CS^(vi) 24.5 ± 1.6 25.4 ± 1.7P1 was dissolved in^(i)TPP;^(ii)NaOH, and added to TPP^(iii)sodium phosphate at pH 6.6, and added to TPP^(iv)CS;^(v)NaOH, and added to CS; and^(vi)sodium phosphate at pH 6.6, and added to CHITOSAN

Example 7

Chitosan (88% deacetylation degree) and glucomannan nanoparticles wereprepared, without incorporating any anionic salt, according to themethod of the invention, with different CS/GM ratios, incorporating P1protein therein. The effectiveness of association of the P1 protein tothe nanoparticles, as well as the load capacity thereof, were measured.TABLE 7 CS/GM Associated Effectiveness of association Load capacity(w/w) protein (%) (%) 6/4.6 P1 37.3 ± 3.8 19.16 ± 1.95 6/13.8 P1 22.4 ±7.1  4.46 ± 1.41

Example 8

Chitosan (88% deacetylation degree) and glucomannan nanoparticles wereprepared, without incorporating any anionic salt, according to themethod of the invention, incorporating P1 protein or insulin therein.The effectiveness of association of the P1 protein and insulin to thenanoparticles, as well as the load capacity thereof, were measured.TABLE 8 CS/GM Associated Effectiveness of association Load capacity(w/w) protein (%) (%) 6/4.6 P1 37.3 ± 3.8 19.16 ± 1.95 6/4.6 Insulin37.5 ± 3.1 23.77 ± 1.78

Example 9

Chitosan (88% deacetylation degree) and glucomannan nanoparticles wereprepared, according to the method of the invention, incorporatingindomethacin or acyclovir therein. The effectiveness of association ofindomethacin and of acyclovir to the nanoparticles, as well as thediameter thereof, were measured. TABLE 9 Effectiveness of associationAssociated drug (%) Mean diameter (nm) Indomethacin 81.81 ± 3.89 619 ±15 Acyclovir 27.36 ± 7.90 304 ± 8 

Example 10

Chitosan (88% deacetylation degree), glucomannan and sodiumtripolyphosphate nanoparticles were prepared, according to the method ofthe invention, with different CS/TPP/GM ratios. The effect thereon ofthe type and the concentration of the cryoprotective agent used in thelyophilization of the nanoparticles on the particle size and zetapotential have been checked (Df: final diameter, Di: initial diameter).(See FIG. 1).

Example 11

Chitosan (88% deacetylation degree), glucomannan and sodiumtripolyphosphate nanoparticles were prepared, according to the method ofthe invention, with different CS/TPP/GM ratios. During their incubationin a phosphate buffer solution at pH 7.4 for 2 hours, the mean diameterof the particles was measured. Theoretical CS/TPP/GM ratios: (

) 311/13.5, (▪) 311/6.7 and (˜) 6/1/23. (See FIG. 2)

Example 12

Chitosan (88% deacetylation degree) and glucomannan nanoparticles wereprepared, according to the method of the invention, with different CS/GMratios. During their incubation in a phosphate buffer solution at pH7.4, the mean diameter of the particles was measured. (˜) CS/GM=6/4.6; (

) CS/GM=6/13.8; (˜) CS/TPP/GM=6/1/4.6. (See FIG. 3)

Example 13

Chitosan (88% deacetylation degree) and glucomannan nanoparticles wereprepared, according to the method of the invention, with different CS/GMratios, incorporating insulin. The release of the insulin in phosphatebuffer at pH 7.4 and 37° C. was measured. (

) CS/TPP/GM=6/1/4.6 and (▪) CS/GM=6/4.6. (See FIG. 4)

Example 14

Chitosan (88% deacetylation degree) and glucomannan nanoparticles wereprepared, according to the method of the invention, with different CS/GMratios, incorporating P1 protein. The release of the insulin inphosphate buffer at pH 7.4 and 37° C. was measured. (

) CS/TPP/GM=6/1/4.6 and (▪) CS/GM=6/4.6. (See FIG. 5)

Example 15

Chitosan (42% and 88% deacetylation degree) and glucomannan nanopartideswere prepared, according to the method of the invention, incorporating¹²⁵I-BSA. The nanoparticles were orally administered to mice, and bloodradioactivity levels were measured after 2, 4, 6 and 24 hours. (See FIG.6)

Example 16

Chitosan and glucomannan nanoparticles were prepared, according to themethod of the invention, incorporating ¹²⁵I-BSA. The nanoparticles wereintraduodenally administered to mice, and blood radioactivity levelswere measured after 0.5, 2 and 24 hours. Chitosan nanoparticles wereused as a control. (See FIG. 7)

Example 17

Chitosan (42% and 88% deacetylation degree) and glucomannannanoparticles were prepared, according to the method of the invention,incorporating ¹²⁵I-BSA. The nanoparticles were orally administered tomice, and radioactivity levels in different tissues were measured after24 hours. (See FIG. 8)

Example 18

Chitosan and glucomannan nanoparticles were prepared, according to themethod of the invention, incorporating ¹²⁵I-BSA. The nanoparticles wereintraduodenally administered to mice, and radioactivity levels indifferent tissues were measured after 24 hours. Chitosan nanoparticleswere used as a control. (See FIG. 9)

1. A method of producing nanoparticles, with a mean diameter equal to or less than 1 m, and incorporating at least one active ingredient, comprising the following steps: a) preparing an aqueous chitosan solution, b) preparing an aqueous glucomannan solution, and c) mixing, under stirring, the solutions of steps a) and b), such that the chitosan and glucomannan nanoparticles are obtained, wherein at least one of the solutions of steps a) and b) contains at least one active ingredient.
 2. A method of producing nanoparticles according to claim 1, wherein the glucomannan solution contains an anionic salt.
 3. A method of producing nanoparticles according to claim 2, wherein the anionic salt is sodium tripolyphosphate.
 4. A method of producing nanoparticles according to claim 3, wherein the sodium tripolyphosphate is at a concentration between 0.1 and 5 mg/mL.
 5. A method of producing nanoparticles according to claim 1 wherein the concentration of the chitosan solution is in the range between 0.5 and 5 mg/mL.
 6. A method of producing nanoparticles according to claim 1 wherein the concentration of the glucomannan solution is in the range between 0.5 and 50 mg/mL.
 7. A method of producing nanoparticles according to claim 1 wherein the ratio between chitosan and glucomannan is between 1:0.1 and 1:100.
 8. A method of producing nanoparticles according to claim 1 wherein the ratio between chitosan and glucomannan is between 1:0.5 and 1:50.
 9. A method of producing nanoparticles according to claim 1 wherein the chitosan solution has a pH between 2 and
 6. 10. A method of producing nanoparticles according to claim 1 wherein the active ingredient is a bioactive macromolecule.
 11. A method of producing nanoparticles according to claim 1 wherein the active ingredient is chosen from the group consisting of insulin, bovine serum albumin and immunogenic proteins.
 12. A method of producing nanoparticles according to claim 1 wherein the active ingredient is a low molecular weight drug.
 13. A method of producing nanoparticles according to claim 1 wherein the active ingredient is c selected from the group consisting of acyclovir and indomethacin.
 14. A method of producing nanoparticles according to claim 1 further comprising an additional step after step c), of lypholizing the nanoparticles.
 15. Nanoparticles with a diameter equal to or less than 1 μm, for the administration of at least one active ingredient, comprising chitosan, glucomannan and at least one active ingredient.
 16. Nanoparticles according to claim 15, produced by of the method according to claim
 1. 17. Nanoparticles according to claim 15 further comprising an anionic salt.
 18. Nanoparticles according to claim 17, wherein the anionic salt is sodium tripolyphosphate.
 19. Nanoparticles according to claim 15, wherein the active ingredient is a bioactive macromolecule.
 20. Nanoparticles according to claim 15 wherein the active ingredient is selected from the group consisting of insulin, bovine serum albumin and immunogenic proteins.
 21. Nanoparticles according to claim 15 wherein the active ingredient is a drug of low molecular weight.
 22. Nanoparticles according to any of claim 15 wherein the active ingredient is selected from the group consisting of acyclovir and indomethacin.
 23. Nanoparticles according to claim 15 wherein the chitosan:glucomannan ratio is between 1:0.02 and 1:100.
 24. Nanoparticles according to claim 15 wherein the chitosan:glucomannan ratio is between 1:0.5 and 1:50.
 25. Nanoparticles according to claim 15 wherein the particles are lyophilized after they are obtained.
 26. A pharmaceutical composition, comprising the nanoparticles according to claim 15 and at least one pharmaceutically acceptable excipient.
 27. A cosmetic composition, comprising the nanoparticles according to claim 15 and at least one cosmetically acceptable excipient.
 28. A pharmaceutical composition, comprising the nanoparticles of claim 25, wherein after the particles are regenerated by addition of water, and at least one pharmaceutically acceptable excipient.
 29. A cosmetic composition, comprising the nanoparticles of claim 25, wherein after the particles are regenerated by addition of water, and at least one cosmetically acceptable excipient. 